Effects of nitrogen forms and calcium amounts on ...

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JOURNAL OF PLANT NUTRITION https://doi.org/10.1080/01904167.2018.1443127

Effects of nitrogen forms and calcium amounts on growth and elemental concentration in Rosa hybrida cv. ‘Vendentta’ S. Mohammad Banijamalia,b, Mohammad Feizian

b

, Hosein Bayata, and Sahar Mirzaeia

a

Ornamental Plants Research Center, Horticultural Sciences Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Mahallat, Iran; bSoil Science Department, Faculty of Agriculture, Lorestan University, Khorramabad, Iran

ABSTRACT

ARTICLE HISTORY

An experiment was conducted to investigate the effects of calcium and nitrogen on quality and quantity of Rosa hybrida in hydroponic culture, using factorial complete randomized design with different levels of ammonium-N (0, 2.5, and 5 mM) and calcium (1.6 and 4.8 mM). The results indicated that ammonium-N concentration of 2 mM increased the number of flowers, length of pedicles, and fresh weight of flower stem per plant. 5 mM of ammonium-N caused a significant decrease in most of the measured characteristics. Increase in calcium concentration enhanced nitrogen, calcium, manganese, and boron; while, potassium, zinc, and copper decreased in the leaf. Flower diameter and fresh weight of flower stems per plant increased significantly. With application of ammonium in the nutrient solution, calcium and potassium concentration in the leaf decreased, whereas phosphorus, zinc, manganese, iron, and boron significantly increased. Therefore, application of 2.5 mM ammonium-N and 4.8 mM calcium are recommended.

Received 8 August 2016 Accepted 2 November 2016 KEYWORDS

Ammonium; calcium; hydroponic; nutrient uptake

Introduction Rose (Rosa hybrida L.) is one of the most important cut flowers in Iran, which currently has approximately 646 hectares assigned to its culture. Mahalat region located in the central part of Iran, is one of the sectors for ornamental plant production, in which 93 hectares of its greenhouse is under rose cultivation (Anonymous, 2012). Vase life of roses is one of the most important characters influencing the quality of rose cut flowers. Due to the short vase life of roses, research has been conducted to reduce losses. Among all the macroelements in nutrient solutions, calcium (Ca) plays an important role in improving the qualitative characteristics of rose cut flowers (Halevy et al., 2001). Several studies have shown the role of calcium in improving the vase life of rose flower (Starkey and Pedersen, 1997; Mortensen, Ottosen and Gislerod, 2001; Torre, Fjeld and Gislerod, 2001). However, studies on how calcium absorption from nutrient solutions may be affected by some nutrient elements can explain the important role of calcium in qualitative characters of rose flower. Calcium absorption by plant is inactive and may be affected by competition with other cations (Marschner, 2012). On the other hand, ammonium-N and potassium (K) are two main cations that are competitors in calcium absorption and may cause a decrease in calcium absorption from the nutrient solutions (Woodson and Boodley, 1982) and lead to a decrease in vase life of the flower; this may increase the waste of this product. Due to lack of information on the suitable levels of ammonium and CONTACT Mohammad Feizian [email protected] Lorestan Province, Tehran Road, Khorramabad, Iran. © 2018 Taylor & Francis Group, LLC

Soil Science Department, Faculty of Agriculture, Lorestan University,

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calcium in nutrient solutions used for hydroponic culture of rose flowers, this research focused on the effects of different forms of ammonium and concentrations of calcium on qualitative and quantitative characteristics of rose (cv. ‘Vendentta’).

Materials and methods Plant material and growth conditions This experiment was carried out in a hydroponic greenhouse at the Ornamental Plants Research Center in Mahallat City, Iran (33o540 3000 N, 50o270 3000 E and 1747 m alt.), starting from spring, for one year. One-year-old shrub rose (Rosa hybrida L.) variety Vendentta, planted in 10 L plastic pots, mixed with fine coconut fiber medium (Coco peat) C Perlite (3–5 mm) in the ratio of 70:30 were used in a factorial complete randomized block design with two factors and four replications. The first factor was ammonium-N levels (0, 2.5, and 5 mM) and the second factor was calcium levels (1.6 and 4.8 mM) in nutrient solution. Total nitrogen (ammonium-N and nitrate-N) in the six nutrient solutions was 10 mM, and the ammonium-N: nitrate-N ratio was 0:100, 25:75, and 50:50%, in the 0, 2.5 and 5 mM ammonium-N levels, respectively. The composition of other macronutrients in the six nutrient solutions was: phosphorus 1, potassium 7.4, and magnesium 2 (mM), and that of the micronutrients was copper 0.32, boron 46, iron 35, manganese 9.14, zinc 0.76, and molybdenum 0.11 (mM). Nutrient elements were obtained from chemical fertilizers of ammonium sulfate [(NH4)2SO4], potassium nitrate (KNO3), calcium nitrate {5[Ca (NO3)2.2H2O].NH4NO3}, potassium nitrate (KNO3), ammonium nitrate (NH4NO3), magnesium nitrate [Mg(NO3)2.6H2O], phosphoric acid (H3PO4), potassium sulfate (K2SO4), magnesium sulfate (MgSO4.7H2O), copper sulfate (CuSO4.5H2O), boric acid (H3BO3), iron chelate (Fe-EDDHA, 6% Fe), manganese sulfate (MnSO4.H2O(, zinc sulfate (ZnSO4.7H2O), and sodium molybdate (Na2MoO4. 2H2O) (Hoagland and Arnon, 1950). In all the nutrient solutions, pH was adjusted between 5.8 and 6.2. In order to adjust pH, 1 molar sulfuric acid solution was used. The electrical conductivity (EC) of all the nutrient solutions was 2.35§ 0.15 (dS/m). Also, washing of pot media was done weekly by water irrigation till the EC was less than that in the nutrient solutions, to inhibit salt accumulation. Each pot contained one plant and four pots were considered for each replication. So, sixteen pots were used for one treatment. Some physical and chemical properties of the medium used in the experiment are shown in Table 1 (Verdonck and Gabriels, 1992; Aliahyaei and Behbahanizadeh, 2002). The light inside the glasshouse was adjusted to 27,000 Lux (with shading), 43,000 Lux (without shading) in the summer, and 28,000 Lux (without shading) in winter. Pots containing rose flowers in the glasshouse were placed on the stages (8 plants/square meter). Hydroponic system that was used in this research work was open form, with dripper irrigation system. Duration of each irrigation was 1.2–1.5 min with 35–40 ml/min dripper, in 4–8 irrigation frequencies, throughout the experiment. For preparation of nutrient solutions, tap water was used and different amounts of nutrient elements that were in the tap water were subtracted from the quantities required (Table 2). The cooling system of the greenhouse was fan and pad. Average daily temperature was 25 C, night temperature was 16 C, and optimum relative humidity was 60%. Petal and leaf samples (from first and second leaves that appeared) were taken in the flowering period (Jones, Wolf and Mills 1991). Control of mites, bugs, and powdery mildew was done during the growth period. Table 1. Physical and chemical properties of used media. Water holding capacity % 73.6

Dry bulk density Fresh bulk density particle density (g/cm3) (g/cm3) (g/cm3) 0.162

0.617

1.28

Total porosity % 87.3

Water Air capacity EC (dS/ pH capacity % % m) 45.5

45.3

1.84 5.90

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Table 2. Tap water utilized for preparing nutritional solutions. pH

SO42¡

Cl¡

CO32¡

7.61

0.545

0.596



HCO3¡ meq/L 2.91

Mg2C

Ca2C

KC

NaC

EC

1.60

1.80



0.765

dS/m 0.405

Vegetative growth parameters During the flowering period, with the appearance of blooms, when sepals had turned down and the flower had become cylindrical, flowers were harvested from spring and different characteristics such as number of flowers per plant, fresh weight of flower stems per plant, length of pedicle (length of last node of stem to flower), flower length, flower diameter, stem length, petal number, and vase life were measured for one year. Fresh weight of flower stems per plant was measured using a digital balance. The height of flower stem and length of pedicle were measured using a ruler, while length and flower diameter were measured using a caliper. In order to determine vase life, cut flowers were transferred to pots containing distilled water and placed in a controlled condition of 20 C temperature, 60% relative humidity, and constant light. When flowers had bent neck or falling petals occurred, the shelf life of flowers was calculated on the basis of the number of days after harvesting. Nutrient elements Oven dried (72 C for 48 hr) samples of leaves and petals were ground separately and passed through a 40-mesh sieve. The ground plant samples were dry-ashed at 500 C for 4 hr; the ashes were dissolved in 10 mL HCl (2N) and the volume was made to 100 mL of distilled water. The concentration of Mg, Ca, Mn, Zn, and Cu were determined by atomic absorption spectrometry (Thermo Elemental Type Solaar 969 MKII England). K concentrations were determined by flame photometry (JENWAY, LTD, PFP7 England) and P concentration in the digest was determined by vanadate molybdate method (yellow method) (Chapman and Pratt, 1961) using spectrophotometry (Spectronic instruments 4001/4 USA). MicroKjeldahl method was used to measure the total N (Bremmer and Malvancy, 1982). Statistical analysis Analysis of variance was performed using the SAS version 9.1.3 software (SAS Inc., Carey NC) and the treatment means were compared by Duncan’s test.

Results Effect of different levels of ammonium-N and calcium on qualitative and quantitative performance of rose flower Analysis of variance indicated that main effects of different levels of ammonium-N on fresh weight of flower stems per plant, number of flowers per plant, vase life, flower diameter, and length of pedicle were statistically significant. Ammonium-N concentration of 2.5 mM caused 21% increase in the number of flowers when compared with the control (without ammonium-N) (Table 3). Vase life also increased by 3.28% with increase in Ammonium-N concentration to 2.5 mM as compared to the control, while 5 mM concentration caused 8.82% decrease, which was statistically significant (Table 3). Length of pedicle, is an effective factor in flower marketing, had 11.8% increase with increases in the ammonium-N concentration to 2.5 mM and, subsequently, decreased with an increase to 5 mM (Table 3). In similar conditions, increases in ammonium-N concentration to 2.5 mM caused an increase in fresh weight of flower stems per plant (18.65%) when compared with the control (Table 3). The 0 and 2.5 mM ammonium-N concentration of the nutrient solution did not show a significant

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Table 3. Main effects of different levels of ammonium-N on some characteristics of rose cut flower. Fresh weight of stem flower per plant (g) 74.46 ab1 88.35a 69.95b

Vase life (Day)

Length pedicle (cm)

Flower diameter (cm)

Flower length (cm)

7.37a 7.62a 6.72b

8.55ab 9.56a 8.12b

2.11a 2.02a 1.93b

3.98a 4.00a 3.96a

Flower number Stem Number per plant length (cm) petals 3.53b 4.28a 3.47b

40.24 a 43.20a 39.35a

10.91a 10.95a 10.38a

Ammonium-N conc. (mM.) 0 2.5 5.0

1

Significant at 5% level of probability by Duncan test.

difference in flower diameter, but when increased to 5 mM caused a significant decrease in flower diameter (Table 3). However, the highest petal number, stem length, and flower length were obtained at 2.5 mM ammonium-N (Table 3). The main effect of calcium concentration on fresh weight of flower stems per plant and flower diameter was significant (Table 4). Increases in calcium concentrations from 1.6 to 4.8 mM caused a significant increase in flower diameter and fresh weight of flower stems per plant (7 and 11.89%, respectively) (Table 4).

Effect of different levels of ammonium-N and calcium on concentration of nutrient elements in rose flower Analysis of variance indicated that main effects of different levels of ammonium-N on the concentration of phosphorus, potassium, calcium, iron, zinc, manganese, and boron in the leaf were significant. Increases in ammonium-N concentration of the nutrient solution from 0 to 2.5 mM caused a significant increase in the concentration of zinc, manganese, and iron in leaf, and then caused a significant decrease in the potassium concentration of leaf (Table 5). Increases in ammonium-N concentration of the nutrient solution from 2.5 to 5 mM caused a significant decrease in calcium concentration of leaf, and a significant increase in phosphorus and boron (Table 5). Analysis of variance showed that the main effects of different levels of ammonium-N on the concentration of zinc, manganese, and copper in the petal were significant. Increases in ammonium-N concentration of the nutrient solution from 0 to 2.5 mM caused significant increase in the concentration of nitrogen, zinc, manganese, and copper in petal (Table 5). The concentration of iron in the petal also increased with increases in ammonium-N concentration of the nutrient solution to 5 mM. An increase from 2.5 to 5 mM caused a significant increase in the concentration of copper in the petal (Table 5). Analysis of variance showed that main effects of different levels of calcium on the concentration of nitrogen, potassium, calcium, zinc, manganese, copper, and boron in the leaf were significant. With an increase in calcium concentration to 4.8 mM, the concentration of nitrogen, calcium, manganese, and boron in the leaf showed a significant increase (Table 6). In contrast, the concentration of potassium, zinc, and copper in the leaf had significant decrease (Table 6). An increase in calcium concentration of nutrient solution caused a significant increase in the concentration of calcium and copper in the petal (Table 6). Table 4. Main effects of different levels of calcium on some characteristics of rose cut flower. Fresh weight of stem flower per plant (g) 73.23b1 81.94a 1

Vase life (Day)

Length pedicle (cm)

Flower diameter (cm)

Flower length (cm)

6.99a 7.48a

8.82a 8.67a

2.01b 2.15a

4.06a 3.89a

Significant at 5% level of probability by Duncan test.

Flower number Stem length Number per plant (cm) petals 3.60a 3.92a

41.3a 40.6a

10.68a 10.75a

Calcium conc. (mM.) 1.6 4.8

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Table 5. Main effects of different levels of ammonium-N on nutritional elements concentration in the leaf and petal of rose. mg/kg B 77.44 b1 87.81b 103.7a 20.63a 18.88 a 30.44 a

%

Cu

Fe

Mn

Zn

Ca

K

P

N

Ammonium-N conc. (mM)

1.000 a 0.625 a 0.813 a 0.526 b 0.938 b 2.625 a

63.70 b 87.75 a 81.38 a 31.63 a 34.81 a 45.63 a

91.31 b 203.90 a 222.60 a 48.06 b 72.69 a 79.25 a

63.13 b 190.7 a 173.0 a 63.13 b 190.7 a 173.0 a

2.849 a 2.66 a 2.438 b 0.461 a 0.441 a 0.412 a

2.748 a 2.386 b 2.310 b 4.156 a 3.219 a 3.579 a

0.472 b 0.486 b 0.539 a 0.250 a 0.274 a 0.240 a

3.96 a 3.67 a 3.71 a 1.57 b 1.92 ab 2.156 a

0 2.5 5.0 0 2.5 5.0

Part of plant Leaf Petal

1

Significant at 5% level of probability by Duncan test.

Discussion Results indicated that replacing 25% of total nitrogen (2.5 mM) of the nutrient solution with ammonium-N (ammonium-N to nitrate-N ratio: 25:75) was beneficial in the improvement of growth and yield of rose flower and increased the fresh weight of flower stems per plant. Similar to the results of the present research, in most researches, applying nitrate-N and ammonium-N together, especially at the ratio of 25% ammonium-N and 75% nitrate-N caused increase in the growth of greenhouse plants, such as rose, in comparison with each of them separately (Hartman, Mills and Jones, 1986; Hohjo et al., 1995). But an increase in ammonium-N ratio in the use of nitrogen to 50% (5 mM) caused a decrease in fresh weight of flower stems production per plant. This decrease might be due to a decrease in pH of the nutrient solution around the root (Errebhi and Wilcox, 1990; Feigin et al., 1986; Magalhaes and Wilcox, 1983) or decrease in calcium absorption by root. A decrease in the pH of the rhizosphere and also calcium absorption is due to the use of ammonium-N fertilizers in the soil media (Cooke, 1967). Herein, results indicated that an increase in calcium of the nutrient solution from 1.6 to 4.8 mM caused a significant increase in fresh weight of flower stems per plant (Table 5) and increased calcium concentration in the leaf and petal of rose (Table 6). Ammonium-N application together with nitrate-N (25% ammonium-N and 75% nitrate-N) caused an increase in the number of flowers in rose (Feigin et al., 1986). It has been reported that the positive effect of ammonium-N application in the ratio of 25% of total nitrogen is due to the lower energy consumption by the plant for absorption and assimilation of ammonium-N in comparison with nitrate-N (Marschner, 2012). This energy saved consists of up to 17% of total carbohydrate resources of the plant (Gutschick, 1981). Increases in ammonium-N: nitrate-N ratio from 25:75 to 50:50 caused a decrease in the number of flowers per plant. More increase in the ammonium-N ratio of the nutrient solution in comparison with nitrate-N was done to provide an acidic condition around the root (pH less than 5) and cause a decrease in root growth and finally reduced plant production (Marschner, 2012). This may be due to harmful effects of free ammonium-N that is stored in plant tissues. If excess ammonium-N absorption and formation cause storage of free ammonium-N in plant tissues, especially in leaves, it can interfere with so many metabolic processes, e.g., photosynthesis (Givan, 1979). The most important index of flower quality in the investigation of qualitative factors is vase life or longevity of the flower. The vase life of flower decreases with an increase in ammonium-N Table 6. Main effects of different levels of ammonium-N on nutritional elements concentration in the leaf and petal of Rose. mg/kg B 84.5 b1 94.8 a 22.6 a 24.0 a 1

%

Cu

Fe

Mn

Zn

Ca

K

P

N

Ammonium-N conc. (mM)

Part of plant

1.00 a 0.60 b 1.10 b 1.60 a

76.0 a 79.2 a 30.2 a 5/44 a

154.0 b 191.2 a 67.1 a 66.2 a

160.0 a 124.5 b 46.8 a 49.3 a

2.29b 3.00 a 0.40 b 0.48 a

2.70a 2.29 b 3.40 a 3.90 a

0.517a 0.482 a 0.250 a 0.260 a

3.62b 3.94 a 1.88 a 1.89 a

1.6 4.8 1.6 4.8

Leaf

Significant at 5% level of probability by Duncan test.

Petal

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concentration from 2.5 to 5 mM (Table 3). Increasing ratio of ammonium-N to nitrate-N in nutrient solution causes interference with calcium absorption by the plant (Table 5) and cell’s actions, which results in acceleration of the aging process in vase life. In this research, increases in ammonium-N concentration of the nutrient solution to 50% of total nitrogen (5 mM) caused a decrease in calcium concentration of the leaf. Researches indicated that calcium plays a role in delaying aging of plant tissues (Ferguson and Drobak, 1988), especially rose petals (Michalczuk et al., 1989). Results showed that with an increase in calcium concentration of the nutrient solution, senescing of the flower was delayed in pot roses (Starkey and Pedersen, 1997; Nielsen and Starkey, 1999). Aging process in rose flowers is affected by proteins, phospholipids of the cell membrane, ethylene, and ATPase activities, which are all affected by calcium nutrition (Faust and Klein, 1974; Ferguson and Drobak, 1988). Increases in calcium concentrations from 1.6 to 4.8 mM in the nutrient solution and increase in calcium concentration in the leaf and petal of rose, especially in the ratio of 25:75 for nitrate-N to ammonium-N (Table 3), caused decrease in speed of aging process with regards to positive effect of calcium on the mentioned points (Table 6). Results indicated that the greatest length of pedicle was obtained at 2.5 mM and in comparison with 5 mM ammonium-N, there was decrease in length of pedicle and flower diameter (Table 3). In the case of calcium, results were different. With an increase in the amount of calcium, flower diameter increased (Table 4). These results are similar to those of Michalczuk et al. (1989) and Torre, Borochov and Halevy (1999), which showed a positive effect of calcium on an increase in flower diameter and fresh weight of flower stems per plant, and also an improvement in the process of flower blooming in vase life. Effect of different ratios of ammonium-N: nitrate-N on the concentration of nutritional elements in the leaf and petal of rose is presented in Table 5 and as shown, nitrogen concentration in the petal significantly increased with increase in the ratio of ammonium-N: nitrate-N. Increase in petal nitrogen can be due to faster ammonium-N absorption in comparison with nitrate-N together with consumption of lesser energy by the plant, which has been proved in applied researches (Britto and Kronzucker, 2002; Lorenzo et al., 2000; Rothstein and Cregg, 2005; Roosta and Schjoerring, 2007; Roosta et al., 2016). Increase in total nitrogen in the condition of synchronous use of ammonium-N together with nitrate-N shows compatibility between ammonium-N and nitrate-N absorption of total nitrogen by rose flower, which is similar to the results of Mengel and Kirkby (2001) and Jampeetong and Brix (2009). Increase in ammonium-N concentration from 2.5 to 5 mM caused a significant increase in phosphorus absorption of the leaf (Table 5), which is due to the positive effect of ammonium-N: nitrate-N ratio in the nutrient solution on absorption of anions such as phosphorus. These results are similar to the findings of Hartman, Mills and Jones (1986), Errebhi and Wilcox (1990) and Lorenzo et al. (2000). When ammonium-N with lower molecular weight (NO3¡) is available in the nutrient solution of the plant, there are less negative charges to balance. Thus, they may take up higher amount of phosphorus to provide anion equivalents and charge balance (Roosta and Schjoerring, 2007; Zhang et al., 2005). Increase in ammonium-N concentration from 0 to 2.5 and 5 mM caused a significant decrease in potassium of leaf (Table 5). Inhibitory effect of ammonium-N on potassium absorption by the plant has been mentioned in the studies of Pill and Lambeth (1977), Fageria (2001), and Rothstein and Cregg (2005). Due to the antagonism relationship between ammonium-N and calcium in absorption by root, with an increase in ammonium-N concentration of the nutrient solution to 5 mM (Table 5), calcium concentration of leaf decreased (Nielsen and Starkey, 1999; Rothstein and Cregg, 2005; Woodson and Boodley, 1982). Increase in ammonium-N: nitrate-N ratio in nutrient solution caused a significant increase in the concentration of zinc and manganese in the leaf and petal, and also iron, boron, and copper in the leaf (Table 5). Although, absorption of micronutrient such as iron, zinc, manganese, and copper is in the form of cation, the form of their absorption by the plant is different for macronutrient cations such as

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potassium and calcium. Increase in ammonium-N: nitrate-N ratio decreased the pH of the nutrient solution around the root; therefore, caused an increase in absorption of low nutrient elements by the plant (Assimakopoulou, 2006; Mengel and Kirkby, 2001; Savvas et al., 2003). In contrast, a decrease in ammonium-N: nitrate-N ratio together with increase in pH around the root, reduced absorption of low nutrient elements (Nikolic and Romheld, 2003). Presence of ammonium-N can increase the concentration of Zn in plants as reported by Assimakopoulou (2006) in spinach, Roosta and Schjoerring (2007) in lettuce, and Clark and et al. (2003) in azalea. Effect of increasing ammonium-N concentration of nutrient solution on boron concentration of leaf was different from other low nutrient elements. In fact, increase in ammonium-N caused an increase in boron concentration of leaf. This increase is due to the accelerating relationship between ammoniumN and anions such as phosphate and borate (Marschner, 2012). Increase in nitrogen in the leaf of plant with increase in calcium concentration of nutrient solution might be due to the role of calcium in increasing pure protein formation in plant (Faust and Klein, 1974). Higher concentration of calcium in nutrient solution caused a significant decrease in potassium concentration of the leaf (Table 6). Inhibitory effects of calcium on potassium absorption in the plant have been mentioned in different works (Hohjo et al., 1995; Kotsiras et al., 2002; Rothstein and Cregg, 2005). With increase in calcium concentration of the nutrient solution, calcium concentration in the leaf and petal of rose also increased (Table 6), which is similar to the results shown by Starkey and Pederse (1997), Nielsen and Starkey (1999), and Mortensen, Ottosen and Gislerod (2001). Significant positive interaction was observed between increased calcium concentration, and manganese and boron concentration in the leaf, and also with copper concentration in the petal. Also, significant negative interaction was observed between increased calcium concentration of nutrient solution, and zinc and copper in the leaf (Table 6), which might be due to some kinds of inhibitory effects between increasing calcium of nutrient solution, and zinc and copper concentrations. Despite this, zinc concentration in the leaf was sufficient for the plant but the amount of copper in the plant was in the level of deficiency (De Kreij et al., 1992). Thus, it is necessary that higher amount of copper is applied in the nutrient solution.

Conclusion and recommendations From the results, increasing ammonium-N: nitrate-N ratio to 25:75 (2.5 mM ammonium-N) in the nutrient solution and applying 4.8 mM of calcium is recommended to improve vegetative characteristics together with production of rose flowers and increase the quality and quantity of rose cut flowers in hydroponic culture.

Acknowledgments The authors thank all the staff in the Ornamental Plants Research Centre (Mahallat-Iran) that helped in this research project.

ORCID Mohammad Feizian

http://orcid.org/0000-0002-3206-4434

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